The lysis of cells in order to extract the nucleic acids or proteins inside it is a crucial unit operation in biomolecular analysis. This paper presents a critical evaluation of the various methods that are available both in the macro and micro scale for cell lysis. Various types of cells, the structure of their membranes are discussed initially. Then, various methods that are currently used to lyse cells in the macroscale are discussed and compared. Subsequently, popular methods for micro scale cell lysis and different microfluidic devices used are detailed with their advantages and disadvantages. Finally, a comparison of different techniques used in microfluidics platform has been presented which will be helpful to select method for a particular application.

A Review on Macroscale and Microscale Cell Lysis Methods

With the fast development of acoustic and multiphase microfluidics in recent years, oscillating bubbles have drawn more-and-more attention due to their great potential in various Lab on a Chip (LOC) applications. Many innovative bubble-based devices have been explored in the past decade. In this article, we first briefly summarize current understanding of the physics of oscillating bubbles, and then critically summarize recent advancements, including some of our original work, on the applications of oscillating bubbles in microfluidic devices. We intend to highlight the advantages of using oscillating bubbles along with the challenges that accompany them. We believe that these emerging studies on microfluidic oscillating bubbles will be revolutionary to the development of next-generation LOC technologies.

Oscillating bubbles: a versatile tool for lab on a chip applications

Bubbles are present in a large variety of emerging applications, from advanced materials to biology and medicine, as either laser-generated or acoustically driven bubbles In these applications, th

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Bubble Dynamics in Soft and Biological Matter

The study of the interaction of bubbles with shock waves and ultrasound is sometimes termed 'acoustic cavitation'. It is of importance in many biomedical applications where sound waves are applied. The use of shock waves and ultrasound in medical treatments is appealing because of their non-invasiveness. In this review, we present a variety of acoustics-bubble interactions, with a focus on shock wave-bubble interaction and bubble cloud phenomena. The dynamics of a single spherically oscillating bubble is rather well understood. However, when there is a nearby surface, the bubble often collapses non-spherically with a high-speed jet. The direction of the jet depends on the 'resistance' of the boundary: the bubble jets towards a rigid boundary, splits up near an elastic boundary, and jets away from a free surface. The presence of a shock wave complicates the bubble dynamics further. We shall discuss both experimental studies using high-speed photography and numerical simulations involving shock wave-bubble interaction. In biomedical applications, instead of a single bubble, often clouds of bubbles appear (consisting of many individual bubbles). The dynamics of such a bubble cloud is even more complex. We shall show some of the phenomena observed in a high-intensity focused ultrasound (HIFU) field. The nonlinear nature of the sound field and the complex inter-bubble interaction in a cloud present challenges to a comprehensive understanding of the physics of the bubble cloud in HIFU. We conclude the article with some comments on the challenges ahead.

https://royalsocietypublishing.org/doi/pdf/10.1098/rsfs.2015.0019

Bubbles with shock waves and ultrasound: a review

We present a programmable, biocompatible technique for dynamically concentrating and patterning particles and cells in a microfluidic device. Since our technique utilizes opto-thermally generated, acoustically activated, surface bubbles, we name it “optoacoustic tweezers”. The optoacoustic tweezers are capable of concentrating particles/cells at any prescribed locations in a microfluidic chamber without the use of permanent structures, rendering it particularly useful for the formation of flexible, complex cell patterns. Additionally, this technique has demonstrated excellent biocompatibility and can be conveniently integrated with other microfluidic units. In our experiments, micro-bubbles were generated by focusing a 405 nm diode laser onto a gold-coated glass chamber. By properly tuning the laser, we demonstrate precise control over the position and size of the generated bubbles. Acoustic waves were then applied to activate the surface bubbles, causing them to oscillate at an optimized frequency. The resulting acoustic radiation force allowed us to locally trap particles/cells, including 15 μm polystyrene beads and HeLa cells, around each bubble. Cell-adhesion tests were also conducted after cell concentrating to confirm the biocompatibility of this technique.

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Optoacoustic tweezers: a programmable, localized cell concentrator based on opto-thermally generated, acoustically activated, surface bubbles

We report on an efficient ultrasound based technique for lysing Escherichia coli and Pichia pastoris with oscillating cavitation bubbles in an integrated microfluidic system. The system consists of a meandering microfluidic channel and four piezoelectric transducers mounted on a glass substrate, with the ultrasound exposure and gas pressure regulated by an automatic control system. Controlled lysis of bacterial and yeast cells expressing green fluorescence protein (GFP) is studied with high-speed photography and fluorescence microscopy, and quantified with real-time polymerase chain reaction (qRT-PCR) and fluorescence intensity. The effectiveness of cell lysis correlates with the duration of ultrasound exposure. Complete lysis can be achieved within one second of ultrasound exposure with a temperature increase of less than 3.3 °C. The rod-shaped E. coli bacteria are disrupted into small fragments in less than 0.4 seconds, while the more robust elliptical P. pastoris yeast cells require around 1.0 second for complete lysis. Fluorescence intensity measurements and qRT-PCR analysis show that functionality of GFP and genomic DNA for downstream analytical assays is maintained.

Leonie Driessen

Papers

Sonolysis of Escherichia coli and Pichia pastoris in microfluidics